Quantitative detection of the Ralstonia solanacearum species complex in soil by qPCR combined with a recombinant internal control strain

利用qPCR结合重组内参菌株对土壤中青枯菌复合种进行定量检测

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作者:Wei Chen,Jun-Wei Zhang,Bi-Xia Qin,Hui-Ting Xie,Zhi Zhang,Xiu-Ze Qiao,Shan-Kui Li,Muhammad Asif,Song Guo,Li-Xian Cui,Pei-Pei Wang,Li-Hong Dong,Qing-Gang Guo,Wen-Jun Jiang,Ping Ma,Zhen-Yuan Xia,Can-Hua Lu,Li-Qun Zhang

Abstract

DNA-based detection and quantification of soil-borne pathogens, such as the Ralstonia solanacearum species complex (RSSC), plays a vital role in risk assessment, but meanwhile, precise quantification is difficult due to the poor purity and yield of the soil DNA retrieved. The internal sample process control (ISPC) strain RsPC we developed solved this problem and significantly improved the accuracy of quantification of RSSC in different soils. ISPC-based quantitative PCR detection is a method especially suitable for the quantitative detection of microbes in complex matrices (such as soil and sludge) containing various PCR inhibitors and for those not easy to lyse (like Gram-positive bacteria, fungi, and thick-wall cells like resting spores). In addition, the use of ISPC strains removes additional workload on the preparation of high-quality template DNA and facilitates the development of high-throughput quantitative detection techniques for soil microbes.

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